CN103765207A - Chromatogram data processing device and processing method - Google Patents

Abstract

A chromatogram data processing device (2) which is an embodiment of the present invention comprises an impurity detection unit (30) which further comprises a differential chromatogram creation unit (25) and an assessment unit (26). The differential chromatogram creation unit (25) obtains a wavelength differential coefficient by deriving an absorbance spectrum in each measurement time for the maximum (or minimum) absorption wavelength of an objective component in the wavelength direction and creates a differential chromatogram which represents a change over time of the wavelength differential coefficient. The assessment unit (26) assesses whether the objective component peak includes an impurity on the basis of the shape of the created differential chromatogram. It is thus possible to assess with high precision whether an impurity is included in the objective component peak without requiring a complicated calculation process.

Description

Chromatography data system and disposal route

Technical field

The present invention relates to process based on chromatography to particularly comprising chromatography data system and the disposal route of carrying out the data that spectral analysis collects with the sample of the separated composition of the cylinder (column) of liquid chromatography (LC) or the sample that imports by Using Flow Injection Analysis.

Background technology

As detecting device, used in the liquid chromatography (LC) of the multi-channel type detecting devices such as PDA (Photo Diode Array) detecting device, the injection of sample of flow phase of take is basic point constantly, for the eluate from cylinder, repeatedly obtain absorbance spectrum, thereby can obtain the three-dimensional chromatographic data with these three dimensions of time, wavelength and absorbance.Figure 13 is the schematic diagram of three-dimensional chromatographic data.By extract the data of specific wavelength among this three-dimensional chromatographic data, thereby can generate the wavelength chromatogram that represents the moment of this specific wavelength and the relation of absorbance.In addition, by extract the data of particular point in time among above-mentioned three-dimensional chromatographic data, thereby can generate the absorbance spectrum that represents the wavelength of this time point and the relation of absorbance.

In this liquid chromatography, while carrying out the quantitative test of known target component, generally, obtain the wavelength chromatogram of the absorbing wavelength corresponding with this target component, the area (or height) that appears at the peak value of the target component origin in this chromatogram is irradiated on lubber-line and calculates quantitative values.

Like this target component is carried out when quantitative, appear at peak value in wavelength chromatogram only origin in target component, be no problem, but peak value not necessarily an origin in single component (target component), often can comprise the beyond thought impurity of analyst.Therefore, in the prior art, carried out peak value purity determination processing always, check a certain peak value occurring in chromatogram be only origin in target component or comprise impurity.

For example, patent documentation 1 discloses by having used peak value purity determination processing method in the chromatogram that the liquid chromatography of multi-channel type detecting device obtains.In the method, by the moment T corresponding with peak value target peak summit in wavelength chromatogram ₀absorbance spectrum be made as S ₀(λ), the absorbance spectrum of any time T is afterwards made as to S (λ),, by following (1) formula, calculates S ₀(λ) with the consistent degree P of S (λ).

[mathematical expression 1]

$P=\frac{\mathrm{\&Sigma;}{S}_0\left(\mathrm{\&lambda;}\right)\&CenterDot;S\left(\mathrm{\&lambda;}\right)}{\sqrt{\mathrm{\&Sigma;}{{\mathrm{<figure-callout\; id="0"\; label="S"\; filenames="HDA0000471949310000051.png,HDA0000471949310000081.png"\; state="\{\{state\}\}">S</figure-callout>}}_0}^2\left(\mathrm{\&lambda;}\right)\&CenterDot;\mathrm{\&Sigma;}{S}^2\left(\mathrm{\&lambda;}\right)}}---\left(1\right)$

And, as shown in figure 14, if if if according to unanimously spend P be 1.0～0.8 be green 0.8～0.6 be yellow 0.6 with next be orange mode, according to peak value summit between the corresponding color of consistent degree P (showing with shade in the drawings), on time-axis direction, cut apart display-object peak value.

If target peak only origin, in target component, as shown in Figure 14 (a), is unanimously spent P higher near peak value summit, away from peak value summit, unanimously spend P just lower, its shape clips the central shaft of peak value and roughly symmetrical.With respect to this, in the situation that the peak value summit of target peak before or after there is impurity peak value (that is, in the situation that target peak comprises impurity), the front or rear consistent degree P on the peak value summit of target peak can reduce.For example, in the example shown in Figure 14 (b), clip peak value summit, and compare in left side, the consistent degree P on right side (on the time by a rear side) is lower.Thus, can be judged as the possibility that comprises impurity near the time range this high.

But in above-mentioned peak value purity decision method of the prior art, if front there is impurity peak value the closelying follow of peak value summit of target peak, due to peak value summit, consistent degree P is nearby less low, therefore cannot correctly judge the existence of impurity.

In addition, in above-mentioned peak value purity decision method, as described in non-patent literature 1, to obtain for judging whether to exist the basis of threshold value of the consistent degree P of impurity peak value, the noise vector that need to be composition by the noise size of for example take under each wavelength is set as parameter.But, go for noise vector and also have the problem that need to carry out following numerous and diverse computing,, in this computing, monitor successively the size of the noise in the provision wavelengths region being detected by multi-channel type detecting device, obtain the standard deviation of the time variation of the noise in this wavelength region may.

In addition, in aforesaid liquid chromatography, in the situation that be that retention time of two and these two compositions is approaching to wanting quantitative target component, sometimes cannot abundant separated origin in the peak value of each target component, as shown in Figure 15 (a), can in chromatogram, there is the peak value overlapping.Now, carried out in the prior art following processing,, as shown in Figure 15 (a), by the tail of decaying (tailing) portion, be front and rear with reading the overlapping peak value vertical segmentation of (reading) portion, calculate respectively the area that is divided into two-part peak value, based on its area value, calculate the quantitative values of each component X, Y.But, like this vertical segmentation in the situation of peak value, owing to can not reflecting each composition effluent profile waveform (that is, the peak value waveform while there are not other compositions) originally, therefore cannot obtain high quantitative accuracy.

In addition,, except vertical segmentation, also know the method for the peak value of next separated each composition of calculation process of for example recording by enforcement patent documentation 2, but this calculation process is extremely miscellaneous, has the problem of processing spended time.In addition, no matter be any method, all as shown in Figure 15 (b) in the situation that the peak value of two target components completely overlapping (side peak value comprises the opposing party's peak value), all cannot separated peak value, therefore cannot carry out quantitatively.

In addition, particularly in the situation that Flow Injection Analysis (the FIA=Flow Injection Analysis) method of not utilizing cylinder (that is to say and do not carry out component separating) is used in the quantitative test of this composition in the sample that comprises single component sometimes.FIA method is to utilize thrower that liquid chromatography uses etc. and to the sample that injects ormal weight in the mobile phase of sending here with certain flow, along with flowing to the method for detecting device importing sample of mobile phase, with identical from the eluate of cylinder outlet, the concentration of target component is varied to roughly chevron shape along with the process of time.While detecting by multi-channel type detecting device the sample importing by this FIA method, resulting data are also the three-dimensional datas with these three dimensions of time, wavelength and absorbance, identical with the data of collecting by liquid chromatography as above in fact.Therefore, " the three-dimensional chromatographic data " of this instructions also comprises the three-dimensional data of collecting by FIA method.

Technical literature formerly

Patent documentation

Patent documentation 1: No. 2936700 communique of patent

Patent documentation 2:JP JP 2006-177980 communique

Non-patent literature

Non-patent literature 1: light husband, " Off ォ ト ダ ィ ォ mono-De ァ レ ィ UV-VIS detecting device SPD-M6A for the HPLC of Island Tianjin ", Island Tianjin Evaluation Theory, the 46th volume, No. 1, in July, 1989, pp.21-28 are deceived in water Kobe Kang Jing, north

Summary of the invention

(problem that invention will solve)

The present invention completes in order to address the above problem, and its 1st object is to provide and is not a kind ofly needing complicated computing just can judge accurately whether target peak comprises chromatography data system and the disposal route of impurity.

In addition, even if the 2nd object of the present invention is to provide, a kind of in the situation that peak value of two target components is overlapping not to be needed complex calculations to process just to carry out the chromatography data system of the quantitative test of these two compositions and disposal route accurately yet.

(for the means of dealing with problems)

Known to reach above-mentioned the 2nd object, two overlapping peak separation of target component need to be become to each target component, if but think that a side in two target components is real target component and the opposing party is undesired impurity, in order to reach above-mentioned the 2nd object, the basic fundamental thought of necessary peak separation also can be used in the situation of reaching the 1st object.

That is, the invention is characterized in of completing in order to reach the above-mentioned the 1st and the 2nd object, processes take in the chromatography data system of the three-dimensional chromatographic data that time, wavelength and absorbance be dimension of collecting for target sample, possesses:

A) differential chromatogram generation unit, based on described three-dimensional chromatographic data, for the wavelength of each time point and the absorbance spectrum of the relation between absorbance that represent in all time ranges or within the scope of the stipulated time, the differential coefficient of very big (or minimum) absorbing wavelength of obtaining the 1st composition in wavelength direction, be wavelength differential coefficient, generate the differential chromatogram that represents that time of the described wavelength differential coefficient in all time ranges or within the scope of the stipulated time changes; With

B) chromatogram waveform processing unit, the waveform based on described differential chromatogram, determines whether quantitative to the 2nd overlapping composition of the peak value of Multiple components or execution and described the 1st composition of overlapping other of the peak value that exists with described the 1st composition 1.

Above-mentioned three-dimensional chromatographic data is typically the data of repeatedly obtaining absorbance spectrum by detecting devices such as multi-channel type detecting devices for the sample that comprises chromatographic cylinder separated composition on time orientation and obtaining.

In addition, can be also the sample that replaces having passed through cylinder, the data that the sample that does not carry out component separating and be just imported into obtained equally by FIA method.

In addition, above-mentioned detecting device can not be multi-channel type detecting device, so long as can obtain the spectrum of waveform shape wider (change slowly), can be also the ultra-violet and visible spectrophotometer that is accompanied by length scanning in order to obtain absorbance spectrum, infrared light spectrophotometer, near infrared light spectrophotometer, fluorospectrophotometer etc.

In addition, above-mentioned chromatography can be a kind of in liquid chromatography, gas chromatography.

Above-mentioned absorbance spectrum represents from the relation between the light wavelength of sample and the absorbance of each wavelength.In this absorbance spectrum, in each material, there is intrinsic very big (or minimum) absorbing wavelength.Greatly (or minimum) absorbing wavelength exists a plurality ofly in most situation for each material, is limited in the situation of provision wavelengths scope and only there will be one.

The acquisition methods of very big (or minimum) absorbing wavelength of the 1st composition is not particularly limited, it can be the method that directly makes operator's incoming wave long value, also can be the method that makes operator's intended target composition, can also be the method for obtaining wavelength value corresponding thereto from database.In addition, also can, according to the automatic detection peak of three-dimensional chromatographic data, by contrasting with database, thereby determine greatly (or minimum) absorbing wavelength.

In the relevant chromatography data system of the present invention, differential chromatogram generation unit is for example read required data from preserve the storage part of the three-dimensional chromatographic data of collecting for target sample, respectively for the wavelength of each time point and the absorbance spectrum of the relation between absorbance that represent in all time ranges or within the scope of the stipulated time, by in wavelength direction, absorbance is carried out to differential in very big (or minimum) absorbing wavelength of the 1st composition, thereby obtain wavelength differential coefficient.Then, in time series, draw the wavelength differential coefficient that each time point in all time ranges or within the scope of the stipulated time is obtained, thereby generate the differential chromatogram of the time variation that has represented wavelength differential coefficient.

As mentioned above, because very big (or minimum) absorbing wavelength is that material is intrinsic, therefore very big (or minimum) absorbing wavelength of heterogeneity is inconsistent conventionally.In addition,, even if occur that once in a while some very big (or minimum) absorbing wavelength are consistent, also can there are mutually different other very big (or minimum) absorbing wavelength.Thus, as long as origin does not comprise other compositions in the peak value of the 1st composition in chromatogram,, at least comprising the absorbance spectrum of each time point of origin in the time range of the peak value of the 1st composition, very big (or minimum) absorbing wavelength of the 1st composition keeps greatly (or minimum).Therefore, the wavelength differential coefficient of this very big (or minimum) absorbing wavelength is put and is all almost 0 at any time, and differential chromatogram is smooth shape.

On the other hand, if origin comprises other compositions in the peak value of the 1st composition,, in the absorbance spectrum of each time point within comprising the time range of these other compositions, very big (or minimum) absorbing wavelength of the 1st composition is subject to the impact of these other compositions and changes.Therefore, in this time range, greatly the wavelength differential coefficient of (or minimum) absorbing wavelength is not 0, and differential chromatogram is not smooth shape.Now, if the amount of other compositions that origin comprises in the peak value of the 1st composition is few, the protuberance occurring in differential chromatogram or recess are just little, if the amount of other compositions that coexist is many, the protuberance occurring in differential chromatogram or recess are just large.That is protuberance or the recess, occurring in this differential chromatogram, the Size-dependent that is to say peak value are in the amount of other compositions that coexist.

If other compositions that coexist are a kind of, the size of regarding the peak value in differential chromatogram as is the amount of this kind of composition.In addition,, even if other compositions that cannot clearly coexist are one or more, also at least can determine and whether have such composition.Therefore, the waveform of chromatogram waveform processing unit based on differential chromatogram, determines whether and has other compositions overlapping with the peak value of the 1st composition, or carries out the quantitative of the 2nd composition overlapping with the peak value of the 1st composition.

If think above-mentioned the 1st composition be target component and overlap 1 to Multiple components be impurity, by above-mentioned chromatogram waveform processing unit, for target component, carry out and to have pure judgement.

That is, the chromatography data system of the 1st mode of the present invention also possesses: c) wavelength chromatogram generation unit, and based on described three-dimensional chromatographic data, about the absorbing wavelength of the 1st composition and generate the wavelength chromatogram of the relation between expression time and absorbance,

Described differential chromatogram generation unit is following unit: based on described three-dimensional chromatographic data, absorbance spectrum for each time point in the time range of the peak value that comprises the target component in described wavelength chromatogram, obtain the wavelength differential coefficient of very big (or minimum) absorbing wavelength of described the 1st composition, generate the differential chromatogram of the time variation that represents this wavelength differential coefficient

Described chromatogram waveform processing unit is following identifying unit: whether the waveform shape based on described differential chromatogram, comprise impurity in the peak value of judgement as described the 1st composition of target component.

At this, as " absorbing wavelength of the 1st composition ", preferably one of very big (or minimum) absorbing wavelength of the 1st composition, but can be also near the wavelength it.In addition, there is a plurality of greatly (or minimum) absorbing wavelength for this composition in the situation that, the wavelength of preferred maximum intensity wherein.In addition, as " very big (or minimum) absorbing wavelength of the 1st composition ", know in advance near the peak value summit of target peak and have impurity peak value, when wanting to judge the number of this impurity, be chosen in the value of in wavelength direction, the absorption spectrum of this impurity having been carried out to differential and having obtained and there is enough big or small very big (or minimum) absorbing wavelength.

In addition, above-mentioned " time range of the peak value that comprises target component " can detect the peak value in wavelength chromatogram and the time range of getting the origin-to-destination of target peak automatically, can be also to input after the front and back of the retention time of the target peak of operator in wavelength chromatogram arrange reasonable time width.

In chromatogram, even in the situation that impurity peak value be present in target peak summit immediately following front (that is to say that the retention time is extremely near), also can in the shape of the differential chromatogram being generated by differential chromatogram generation unit, reflect there is inclusion-free.Therefore,, according to the chromatography data system of the 1st mode, can judge accurately whether target peak comprises impurity.

In addition, according to above-mentioned formation, not in all time ranges but only in " time range of the peak value that comprises target component ", to generate differential chromatogram, thereby judged in this time range and had inclusion-free, therefore not only can more effectively judge whether target peak comprises impurity, can also further shorten the required time of judging.

In addition, in above-mentioned the 1st mode, above-mentioned identifying unit is as long as specifically pass through to judge that whether differential chromatogram is smooth, thereby whether the peak value of judgement target component comprises impurity.

In addition, if consider, the above-mentioned the 1st and the 2nd composition is all known target component, by chromatogram waveform processing unit, carries out the quantitative of two target components.

That is, in the chromatography data system of the 2nd mode of the present invention,

Described differential chromatogram generation unit is based on described three-dimensional chromatographic data, absorbance spectrum for each time point in all time ranges or within the scope of the stipulated time, obtain the wavelength differential coefficient as very big (or minimum) absorbing wavelength of described the 1st composition of the 1st target component, generate the differential chromatogram of the time variation that represents this wavelength differential coefficient, and obtain the wavelength differential coefficient as very big (or minimum) absorbing wavelength of described the 2nd composition of the 2nd target component, generate the differential chromatogram of the time variation that represents this wavelength differential coefficient

The peak value occurring in the differential chromatogram of described chromatogram waveform processing unit very big (or minimum) absorbing wavelength based at described the 1st target component carries out quantitatively described the 2nd target component, and the peak value occurring in the differential chromatogram of very big (or minimum) absorbing wavelength based at described the 2nd target component carries out quantitatively described the 1st target component.

The concentration of size reflection the 2nd target component of the peak value occurring in the differential chromatogram of very big (or minimum) absorbing wavelength of the 1st target component as mentioned above.For example, if the concentration of the 2nd target component is 0, in the differential chromatogram of very big (or minimum) absorbing wavelength of the 1st target component, there will not be peak value, be smooth shape.The concentration of size reflection the 1st target component of the peak value occurring in the differential chromatogram of very big (or minimum) absorbing wavelength of the 2nd target component on the contrary.For example, if the concentration of the 1st target component is 0, in the differential chromatogram of very big (or minimum) absorbing wavelength of the 2nd target component, there will not be peak value, be smooth shape.That is,, if pay close attention to the peak value occurring in the differential chromatogram of very big (or minimum) absorbing wavelength of a target component, can get rid of the impact of this target component and carry out the quantitative of another target component.

At this, very big (or minimum) absorbing wavelength of the 2nd composition needs only by the identical method of acquisition methods of very big (or minimum) absorbing wavelength of the 1st composition with above-mentioned and obtains.That is, owing to wanting, quantitative target component is known, can be therefore the method for these wavelength value of directly being inputted by operator, can be also the method that makes operator's intended target composition, can also be the method for obtaining wavelength value corresponding thereto from database.

In the chromatography data system of above-mentioned the 2nd mode, with used common chromatogram on the quantitative Treatment of the peak value that occurs identical, as long as carry out the quantitative Treatment of the peak value based on occurring in differential chromatogram.That is,, in the chromatography data system of the 2nd mode, the structure of described chromatogram waveform processing unit comprises:

Inspection amount information memory cell, for described the 1st target component and the 2nd target component, storage list is shown in the area of the peak value occurring in differential chromatogram or the inspection amount information of the relation between height and constituent concentration respectively;

Peak information computing unit, calculate the three-dimensional chromatographic data based on for target sample and generate respectively, appear at the area of the peak value in differential chromatogram or highly in very big (or minimum) absorbing wavelength of described the 1st target component and very big (or minimum) absorbing wavelength of described the 2nd target component; With

Quantitative values computing unit, contrasts the area of the peak value being calculated by this peak information computing unit or height with described inspection amount information, obtain the quantitative values of each target component.

Preferably to comprising respectively actual analysis of sample (that is to say standard model) of the 1st known target component of concentration, the 2nd target component, thereby generate inspection amount information, for example lubber-line of storing in inspection amount information memory cell.This also with used common chromatogram on the quantitative Treatment of the peak value that occurs identical.

The generation of above-mentioned differential chromatogram is extremely simply to process, as long as utilized the quantitative Treatment of the inspection amount information generating in advance for differential chromatogram, just can within the very short time, carry out the calculating of quantitative values.Therefore, according to this, form, even if origin is overlapping in the peak value of two target components, also can to each target component, carry out quantitatively rapidly.In addition, appear at the shape of the peak value in differential chromatogram similar in appearance to the desirable profile waveform of having got rid of the impact of overlapping composition, therefore can realize high-precision quantitative.

In addition, the chromatographic data disposal methods involved in the present invention completing in order to reach the above-mentioned the 1st and the 2nd object is collected for target sample take the three-dimensional chromatographic data that time, wavelength and absorbance be dimension, this chromatographic data disposal route is characterised in that, comprising:

A) differential chromatogram generates step, based on described three-dimensional chromatographic data, for the wavelength of each time point and the absorbance spectrum of the relation between absorbance that represent in all time ranges or within the scope of the stipulated time, the differential coefficient of very big (or minimum) absorbing wavelength of obtaining the 1st composition in wavelength direction, be wavelength differential coefficient, generate the differential chromatogram that represents that time of the described wavelength differential coefficient in all time ranges or within the scope of the stipulated time changes; With

B) chromatogram waveform processing step, the waveform based on described differential chromatogram, determines whether quantitative to the 2nd overlapping composition of the peak value of Multiple components or execution and described the 1st composition of overlapping other of the peak value that exists with described the 1st composition 1.

The 1st mode of the chromatographic data disposal route that in addition, the present invention is relevant also comprises:

C) wavelength chromatogram generates step, based on described three-dimensional chromatographic data, generates the wavelength chromatogram of the relation between expression time and absorbance about the absorbing wavelength of the 1st composition,

In described differential chromatogram, generate in step, based on described three-dimensional chromatographic data, absorbance spectrum for each time point in the time range of the peak value that comprises the target component in described wavelength chromatogram, obtain the wavelength differential coefficient of very big (or minimum) absorbing wavelength of described the 1st composition, generate the differential chromatogram of the time variation that represents this wavelength differential coefficient

In described chromatogram waveform processing step, whether the waveform shape based on described differential chromatogram, comprise impurity in the peak value of judgement as described the 1st composition of target component.

And then in addition, the 2nd mode of the chromatographic data disposal route that the present invention is relevant,

In described differential chromatogram, generate in step, based on described three-dimensional chromatographic data, absorbance spectrum for each time point in all time ranges or within the scope of the stipulated time, obtain the wavelength differential coefficient as very big (or minimum) absorbing wavelength of described the 1st composition of the 1st target component, generate the differential chromatogram of the time variation that represents this wavelength differential coefficient, and obtain the wavelength differential coefficient as very big (or minimum) absorbing wavelength of described the 2nd composition of the 2nd target component, generate the differential chromatogram of the time variation that represents this wavelength differential coefficient

In described chromatogram waveform processing step, the peak value occurring in very big absorbing wavelength based at described the 1st target component or the differential chromatogram of minimum absorbing wavelength carries out quantitatively described the 2nd target component, and the peak value occurring in the differential chromatogram of very big (or minimum) absorbing wavelength based at described the 2nd target component carries out quantitatively described the 1st target component.

(invention effect)

According to chromatography data system of the present invention and disposal route, even if be present in the situation of closelying follow front grade, can being missed on the summit of target peak for impurity peak value in chromatogram in the peak value purity of prior art is judged, also can judge accurately whether target peak comprises impurity.In addition, be different from above-mentioned peak value purity determination processing of the prior art, do not need to set noise vector as parameter, therefore can judge whether target peak comprises impurity by fairly simple computing.

In addition, relevant chromatography data system and the disposal route according to the present invention, even in the peak value of two target components, an overlapping or side is comprised in the situation in the opposing party completely for origin in chromatogram, also can carry out based on origin in the profile waveform of each target component high-precision quantitatively.In addition, this quantitative in order to carry out, as long as compare simple calculations, process, even if therefore use the low personal computer of price, also can promptly carry out quantitatively.

Accompanying drawing explanation

Fig. 1 is the schematic configuration diagram of liquid chromatography that possesses the chromatography data system of one embodiment of the invention.

Fig. 2 means the greatly figure of an example of the peak value of (or minimum) absorbing wavelength chromatogram.

Fig. 3 means the figure of an example of the absorbance spectrum of each Measuring Time point.

Fig. 4 means the figure of an example of the absorbance spectrum of target component and impurity.

Fig. 5 means the figure of an example of differential chromatogram.

Fig. 6 means the process flow diagram of the peak value purity determination processing action in the chromatography data system of the present embodiment.

Fig. 7 is the schematic configuration diagram of liquid chromatography that possesses the chromatography data system of other embodiment of the present invention.

Fig. 8 is the figure of an example of the absorbance spectrum of the principle judged of explanation two kinds of composition peak separation of the present invention and peak value purity.

Fig. 9 means the figure of the differential smoothing of the absorbance spectrum based on shown in Fig. 8.

Figure 10 means the figure of two kinds of composition mixing peak values in chromatogram.

Figure 11 means the figure of the differential chromatogram of the differential smoothing based on shown in Fig. 9.

Figure 12 is the schematic configuration diagram of liquid chromatography that possesses the chromatography data system of other embodiment of the present invention.

Figure 13 means according to the schematic diagram of very big (or minimum) absorbing wavelength chromatogram of three-dimensional chromatographic data and the generation of this three-dimensional chromatographic data.

Figure 14 is the display case of the result that obtains by peak value purity determination processing method of the prior art, is (a) example of the peak value that do not comprise impurity, is (b) example of the peak value that comprises impurity.

Figure 15 means the figure of an example of the chromatogram of the state that separated peak value insufficient and two kinds of compositions is overlapping.

Embodiment

[the principle explanation that two kinds of composition peak separation of the present invention and peak value purity are judged]

First, with reference to accompanying drawing 8～Figure 11, illustrate for two kinds of composition peak separation of the present invention of the three-dimensional chromatographic data execution shown in Figure 13 and the principle that peak value purity is judged.

Now, consider that two compositions that consist of x, y are included in the situation in sample.Fig. 8 means the figure of an example of this composition x and y absorbance spectrum separately.As shown in the drawing, general, very big (or minimum) absorbing wavelength corresponding from summit absorbance peak value (greatly (or minimum) point) each material is different.

Fig. 9 is by the absorbance spectrum shown in Fig. 8 being carried out in wavelength direction to the differential smoothing that differential is obtained.In wavelength direction, one section of its differential coefficient that curve rises be on the occasion of, one section of its differential coefficient that curve declines is negative value, on the summit of absorbance peak value and the bottom differential coefficient of trough portion be 0.As shown in Figure 9, the wavelength that is 0 (being " 0 " of differential coefficient from the situation on the occasion of becoming negative value) by differential coefficient in the differential smoothing at composition x is made as λ x, and the wavelength that is 0 (being " 0 " of differential coefficient from the situation on the occasion of becoming negative value equally) by differential coefficient in the differential smoothing at composition y is made as λ y.That is to say, at this, λ x is the very big absorbing wavelength of composition x, and λ y is the very big absorbing wavelength of composition y.

Figure 10 represents that in chromatogram an example and the overlapping state of these peak value profiles of peak value profile are the figure of unsegregated mixing peak value separately for composition x and composition y.Composition x very approaching with retention time composition y, according to mixing peak value, predict that the peak value profile of each composition x, y is very difficult.

The absorbance spectrum of hypothesis composition x is that x (λ), peak value profile are a (t) now, the absorbance spectrum of same hypothesis composition y is that y (λ), peak value profile are b (t), two kinds of three-dimensional chromatogram S (t, λ) that become under subsystem of composition x and composition y (that is to say in chromatogram upward peak overlapping) can have been represented to flow out by following (2) formula.

S(t，λ)＝a(t)x(λ)+b(t)y(λ) …(2)

If (2) formula is carried out to partial differential with wavelength X, as following (3) formula.

$\&PartialD;S(t,\mathrm{\&lambda;})/\&PartialD;\mathrm{\&lambda;}=a\left(t\right)x^{\&prime;}\left(\mathrm{\&lambda;}\right)+b\left(t\right)y^{\&prime;}\left(\mathrm{\&lambda;}\right)---\left(3\right)$

If the wavelength X x that the differential coefficient in (3) formula in the differential smoothing of substitution composition x is 0, due to x ' (λ x)=0, so relation is as follows.

$\&PartialD;S(t,\mathrm{\&lambda;x})/\&PartialD;\mathrm{\&lambda;}=b\left(t\right)y^{\&prime;}\left(\mathrm{\&lambda;x}\right)---\left(4\right)$

Equally, if the wavelength X y that the differential coefficient in the differential smoothing of substitution composition y is 0 in (3) formula, due to y ' (λ y)=0, so relation is as follows.

$\&PartialD;S(t,\mathrm{\&lambda;y})/\&PartialD;\mathrm{\&lambda;}=a\left(t\right)x^{\&prime;}\left(\mathrm{\&lambda;y}\right)---\left(5\right)$

(a) of Figure 11 drawn the result of (4) formulas on time orientation, (b) on time orientation, drawn equally the result of (5) formulas.That is to say, Figure 11 (a) is the differential chromatogram of wavelength X x, and Figure 11 (b) is the differential chromatogram of wavelength X y.Known according to (4) formula, in the differential chromatogram of wavelength X x, only there is the peak value profile b (t) of composition y.In addition, known according to (5) formula, in the differential chromatogram of wavelength X y, only there is the peak value profile a (t) of composition x.The area of these peak value profiles a (t), b (t) and highly depend on the concentration of composition separately.In addition, about the above-mentioned explanation of Fig. 9～Figure 11, be the explanation while having utilized very big absorbing wavelength λ x, the λ y of composition x, y, but replace very big absorbing wavelength, also can utilize the minimum absorbing wavelength of composition x, y.

As previously discussed, even known, flowing out under the situation of these two compositions of x, y, the differential smoothing of the differential coefficient of very big (or minimum) the absorbing wavelength λ x drawn x composition if use on time orientation in wavelength direction, can only isolate y becomes to assign to carry out quantitatively, the differential smoothing of the differential coefficient of very big (or minimum) the absorbing wavelength λ y drawn y composition if use on time orientation in wavelength direction, only separated x becomes to assign to carry out quantitative.

In addition, as shown in Figure 11 (b), the peak value profile of the composition x greatly occurring in the differential chromatogram of (or minimum) absorbing wavelength λ y is negative peak, is carrying out when quantitative as long as positive-negative polarity is overturn.

Pay close attention to now Figure 11 (a), if there is not peak value in this differential chromatogram, if that is to say, differential coefficient is 0 always, this means and does not have composition y.That is,, if whether produced peak value in the differential chromatogram of very big (or minimum) absorbing wavelength λ x of judgement composition x, just can judge that composition y has zero lap.As long as known, carry out this judgement, just, without very big (or minimum) the absorbing wavelength λ y that knows in advance composition y, composition y itself can be unknown composition.Further, as long as judge whether the peak value of the chromatogram of a certain principal component has comprised other compositions, these other compositions may not be for a kind of, can as impurity, process using 1 to Multiple components unification.

That is, in the three-dimensional chromatogram of target component x, be a (t) x (λ), wherein as impurity, sneaked into other 1 to the situation of Multiple components, can represent three-dimensional chromatogram S (t, λ) by following (6) formula.S(t，λ)＝a(t)x(λ)+b(t)y(λ)+c(t)z(λ)+… …(6)

With wavelength X, this (6) formula is carried out to partial differential, the wavelength X x that the differential smoothing x ' of substitution target component x value (λ) is 0, becomes following (7) formula.

$\&PartialD;S(t,\mathrm{\&lambda;x})/\&PartialD;\mathrm{\&lambda;}=b\left(t\right)y^{\&prime;}\left(\mathrm{\&lambda;x}\right)+c\left(t\right)z^{\&prime;}\left(\mathrm{\&lambda;x}\right)+\&CenterDot;\&CenterDot;\&CenterDot;---\left(7\right)$

Should (7) formula be the differential chromatogram of very big (or minimum) absorbing wavelength λ x of target component x, known origin be removed in the peak value of target component x, has only occurred the peak value of impurity.

Thus, by the identical principle of the peak separation of two kinds of compositions with above-mentioned, known can also being determined with without being blended into the impurity in target component.

[structure of the chromatography data system of the 1st embodiment of the present invention and action]

One embodiment (the 1st embodiment) of chromatography data system of the present invention then, is described with reference to Fig. 1～Fig. 6.The 1st embodiment carries out the judgement of peak value purity based on above-mentioned principle.Fig. 1 is the schematic configuration diagram of the liquid chromatography system of the chromatography data system (being designated hereinafter simply as " data processing equipment ") that possesses the present embodiment.

For collecting the LC portion 1 of three-dimensional chromatographic data, liquid-feeding pump 12 attracts mobile phase from mobile phase container 11, with constant flow, gives sample injection unit 13.Sample injection unit 13 is injected sample in the moment of regulation in mobile phase.Sample is sent in cylinder 14 by mobile phase, and each composition during through cylinder 14 in sample is separated on time orientation, from cylinder 14, flows out.

In the outlet of cylinder 14, as the detecting device of the sample composition in the eluate for detection of from cylinder 14 and be provided with a kind of PDA detecting device 15 that belongs to multi-channel type detecting device.PDA detecting device 15 irradiates the light from not shown light source to efflux, and the light that has made to see through eluate by wavelength disperses, and by PDA linear transducer, almost detects the light intensity of each wavelength simultaneously.The detection signal repeatedly being obtained by this PDA detecting device 15 is output to data processing equipment 2 as three-dimensional chromatographic data after being transformed into digital signal by A/D transducer 16.

Data processing equipment 2 comprises: three-dimensional data storage portion 21, for preserving from the three-dimensional chromatographic data of A/D transducer 16 outputs; Wavelength chromatogram generating unit 22, represents according to three-dimensional chromatographic data generation the wavelength chromatogram that the time of the absorbance of provision wavelengths changes; Peak value test section 23, detects the peak value in this wavelength chromatogram; With impurity test section 24, among the peak value detecting, detecting the impurity in the target peak of operator's appointment.In addition, in the present embodiment, wavelength chromatogram generating unit 22 generates very big (or minimum) the absorbing wavelength λ that represents target component _s0very big (or minimum) absorbing wavelength chromatogram of changing of time of absorbance.

Impurity test section 24 comprises as functional module: differential chromatogram generating unit 25, very big (or minimum) the absorbing wavelength λ based on three-dimensional chromatographic data and target component _s0, generate differential chromatogram; With detection unit 26, the shape based on differential chromatogram, judge in target peak and have inclusion-free.The action of these each several parts will be narrated in the back.

Display part 3 is for showing the various information such as very big (or minimum) absorbing wavelength chromatogram, absorbance spectrum, differential chromatogram and result of determination.In order to make operator input also very big (or minimum) absorbing wavelength λ of target setting composition _s0the information required etc. data processing etc., operate operating portion 4.

In addition, the partial function of data processing equipment 2 or repertoire can be reached by the special use control/process software of carrying out personal computer or be arranged in workstation.In addition, display part 3 is general LCD monitor etc., and operating portion 4 can be as indicators such as the keyboard of the standard facility of personal computer or workstation or mouse etc.

Then,, with reference to the process flow diagram of Fig. 6, the distinctive data processing action in the liquid chromatography system of the 1st embodiment is described.

First, the chromatography analysis of carrying out target sample in LC portion 1, from PDA detecting device 15, to 21 outputs of three-dimensional data storage portion, represent the three-dimensional chromatographic data (with reference to Figure 13 (a)) that the time of the absorbance spectrum of provision wavelengths scope changes, and be kept in this three-dimensional data storage portion 21 (step S1).

Then very big (or minimum) absorbing wavelength λ of the target component (for example wanting quantitative composition) that, operator comprises by operating portion 4 input samples _s0wavelength value (step S2).Accept after this input wavelength chromatogram generating unit 22 very big (or minimum) absorbing wavelength λ based on inputted _s0and be kept at the three-dimensional chromatographic data in three-dimensional data storage portion 21, be created on transverse axis and drawn the time, at the longitudinal axis, drawn greatly (or minimum) absorbing wavelength λ _s0very big (or minimum) absorbing wavelength chromatogram (step S3) of absorbance.Figure 13 (b) represents an example of very big (or minimum) absorbing wavelength chromatogram that the three-dimensional chromatographic data based on shown in Figure 13 (a) generates.

Peak value test section 23 is investigated successively the tilt quantity of the curve of very big (or minimum) absorbing wavelength chromatogram being generated by wavelength chromatogram generating unit 22 on time orientation, as shown in Figure 2, in this tilt quantity, becomes the starting point T that setting is judged as YES peak value when above _s, in tilt quantity from being judged as YES peak value summit T while further becoming negative value on the occasion of becoming 0 ₀, at the absolute value of tilt quantity, become the terminal T that setting is judged as YES peak value when following _e, detection peak (step S4) thus.In Fig. 2, only show a peak value, but the in the situation that of including a plurality of composition in sample, conventionally can detect a plurality of peak values.The information of the peak value detecting is displayed on the picture of display part 3, and operator selects origin in the target peak (step S5) of target component from these a plurality of peak values by operating portion 4.

If target peak is selected, differential chromatogram generating unit 25 is obtained the starting point T of target peak from three-dimensional data storage portion 21 _sto terminal T _etime range in absorbance spectrum, for each absorbance spectrum, respectively in wavelength direction to very big (or minimum) absorbing wavelength λ _s0absorbance carry out differential, thereby obtain wavelength differential coefficient (step S6).Then, be created on that transverse axis draws the time and the differential chromatogram (step S7) that drawn the wavelength differential coefficient calculating at the longitudinal axis.Fig. 5 represents an example of differential chromatogram.

Differential chromatogram based on being generated by differential chromatogram generating unit 25, detection unit 26 is implemented the following processing of the principle based on above-mentioned, thereby at the starting point T of target peak _sto terminal T _etime range in be determined with inclusion-free.

Fig. 4 is the T of point sometime schematically showing in chromatography analysis _uthe pattern ((1) in Fig. 4) of absorbance spectrum of target component and the figure of the pattern ((2) in Fig. 4) of the absorbance spectrum of impurity.Like this in the situation that the absorbing wavelength territory of target component and the absorbing wavelength territory of impurity are overlapped, at above-mentioned time point T _uthe pattern of the actual absorbance spectrum obtaining has been added the pattern of absorbance spectrum of target component and the pattern of the absorbance spectrum of impurity and the pattern ((3) in Fig. 4) that obtains.Therefore, very big (or minimum) position of this absorbance spectrum and very big (or minimum) absorbing wavelength λ of target component _s0and inconsistent.

In target peak only origin is in target component in the situation that, as shown in Figure 3, at the starting point T of target peak _sto terminal T _eduring in any time point on, due to very big (or minimum) absorbing wavelength λ of target component _s0with very big (or minimum) position consistency of absorbance spectrum under each time point, therefore very big (or minimum) absorbing wavelength λ _s0wavelength differential coefficient become 0.Therefore, the starting point T of target peak _sto terminal T _etime range in differential chromatogram become the smooth state that has only comprised inevitable noise representing with solid line in Fig. 5.On the other hand, in the situation that target peak includes impurity, as shown in Figure 4, due to very big (or minimum) absorbing wavelength λ of target component _s0inconsistent with very big (or minimum) position of the absorbance spectrum of each time point, therefore very big (or minimum) absorbing wavelength λ _s0wavelength differential coefficient at time range T _s～T _einside become the value beyond 0.Therefore, differential chromatogram, as shown in the dotted line in Fig. 5, can not become smooth state within comprising the time domain of impurity.

Therefore the starting point T that, detection unit 26 is judged in target peak _sto terminal T _etime range in differential chromatogram whether smooth (step S8).If in this time range, differential chromatogram is smooth (being "Yes" in the step S8 of Fig. 6), detection unit 26 be judged to be target peak in this time range, do not comprise impurity, be only origin in the peak value (step S9) of target component.On the other hand, if in above-mentioned time range differential chromatogram uneven (be "No" in step S8), detection unit 26 judgement target peak comprise impurity (step S10) in this time range.The result of determination obtaining like this to operator notification by display part 3 (step S11).

Judge that whether differential chromatogram is smooth for example by determining whether average N times or peak value more than regulation peak area of the noise intensity that has baseline.In addition, can be also the decision method beyond this.

As previously discussed, in the data processing equipment 2 of the 1st embodiment, by the starting point T of the target peak in very big (or minimum) absorbing wavelength chromatogram _sto terminal T _ein time range during this time, judge that whether differential chromatogram is smooth, thereby judge whether target peak comprises impurity.Even in the situation that closelying follow of the peak value summit of target peak front comprises impurity, as long as very big (or minimum) absorbing wavelength λ of target component _s0departed from a little greatly (or minimum), just can have inclusion-free in reflecting in shape of differential chromatogram.Therefore, according to the data processing equipment 2 of the 1st embodiment, compare with peak value purity decision method of the prior art, can access the result of determination of very high degree of precision.

In addition, in the data processing equipment 2 of the 1st embodiment, be different from above-mentioned prior art, owing to not needing to set as parameter the noise vector that the noise contribution by each wavelength forms, therefore can be by carrying out the judgement of peak value purity than the simpler computing of prior art.

In addition, in the data processing equipment 2 of the 1st embodiment, not in the gamut of Measuring Time, to generate differential chromatogram, but at the starting point T of target peak _sto terminal T _etime range in generate differential chromatogram, therefore can more effectively judge whether target peak comprises impurity, can within the shorter time, finish judgement.

In addition, in the above description, operator has selected target peak by operating portion 4, but can not be also now to select a peak value but select a plurality of peak values as target peak.Now, as long as implement above-mentioned impurity by selected each target peak, detect.In addition, also can be not limited to the quantity of the peak value detecting, but preset the detection that all peak values that detect are in pairs implemented impurity.Now, in above-mentioned process flow diagram, omit the processing of step S5.

In addition,, in the 1st embodiment, in step S2, operator has inputted very big (or minimum) absorbing wavelength λ of target component _s0wavelength value, but also can be by the title of operator's intended target composition and structural formula etc., and from database, obtain wavelength value corresponding thereto.Or, also can omit the input itself that operator carries out, for example, according to the automatic detection peak of three-dimensional chromatographic data (three-dimensional peak value), contrast this result and database, thereby determine greatly (or minimum) absorbing wavelength.In the situation that target component has a plurality of greatly (or minimum) absorbing wavelength, as long as use one of them.

And then in addition, setting about the time range that comprises target peak, also can be pre-entered by operating portion 4 time range that is provided with reasonable time width around retention time of the target peak of operator in wavelength chromatogram, thereby obtain the time T corresponding with the starting point of target peak _sand the time T corresponding with terminal _e.

And then in addition, also can on the picture of display part 3, show that wavelength chromatogram as shown in Figure 2, operator are observed picture and, by operating portion 4, specify the time T corresponding with the starting point of target peak _sand the time T corresponding with terminal _e.

If this structure can operator directly be inputted above-mentioned time range in step S5, or in wavelength chromatogram, by clicking operation etc., specify the position of starting point and terminal, thereby determine the starting point T of target peak _sto terminal T _etime range.

And then in addition, in the 1st embodiment, wavelength chromatogram generating unit 22 has generated the wavelength chromatogram of very big (or minimum) absorbing wavelength of target component, but can be also near the wavelength chromatogram of the wavelength of very big (or minimum) absorbing wavelength of target component.In addition,, in the situation that there is a plurality of greatly (or minimum) absorbing wavelength, the wavelength of maximum intensity is wherein selected in general expectation.In addition, very big (or minimum) absorbing wavelength as the 1st composition, knownly in advance near the peak value summit of target peak, there is impurity peak value, when wanting to judge the number of this impurity, be chosen in wavelength direction the value that the absorption spectrum of this impurity has been carried out to differential and there is enough big or small very big (or minimum) absorbing wavelength.

[structure of the chromatography data system of the variation of the 1st embodiment and action]

The data processing equipment of the variation of the 1st embodiment then, is described by Fig. 7.As shown in Figure 7, the data processing equipment of this variation has appended greatly (or minimum) absorbing wavelength acquisition unit 27 in the data processing equipment of above-mentioned the 1st embodiment.

The data processing equipment of this variation has following structure: at very big (or minimum) absorbing wavelength λ of target component _s0be not known and also do not possess for obtaining in the situation of database of this wavelength, the sample of the standard specification that actual measurement comprises target component obtains its greatly (or minimum) absorbing wavelength λ _s0, and the processing for rear class by this value.

First, by LC portion 1, measure the standard model of the standard specification that comprises target component, thereby obtain three-dimensional chromatographic data.The three-dimensional chromatographic data obtaining is stored in three-dimensional data storage portion 21.If operator selects to there will be origin in the suitable wavelength of the peak value of target component, wavelength chromatogram generating unit 22 is read suitable three-dimensional chromatographic data from three-dimensional data storage portion 21, generates the wavelength chromatogram of selected wavelength.Peak value test section 23, according to this wavelength chromatogram, by processing detection peak same as the previously described embodiments, obtains the starting point T with peak value _s, summit T ₀and terminal T _ethe corresponding time.Operator selects origin in the peak value (now, conventionally only can detect a peak value) of the standard specification of target component from the peak value detecting by operating portion 4.

Then, greatly (or minimum) absorbing wavelength acquisition unit 27 is read and is measured the standard model comprise target component and the three-dimensional chromatographic data obtaining from three-dimensional data storage portion 21, and obtains the moment T corresponding with the summit of the peak value of operator's appointment from peak value test section 23 ₀.Then, greatly (or minimum) absorbing wavelength acquisition unit 27 in wavelength direction successively to moment T ₀under the absorbance of absorbance spectrum carry out differential, obtain the wavelength differential coefficient of each wavelength.

Then, obtain wavelength differential coefficient and be 0 wavelength, very big (or minimum) absorbing wavelength λ of the standard specification using it as target component _s0obtain.Very big (or minimum) absorbing wavelength λ of the standard specification of obtaining _s0in the peak value purity determination processing of the sample of measuring after being used to.Getting a plurality of greatly (or minimum) absorbing wavelength λ _s0situation under, as long as operator selects to be judged to be most suitable very big (or minimum) absorbing wavelength λ by operating portion 4 _s0.Thus, if determined very big (or minimum) absorbing wavelength λ of target component _s0, the peak value purity determination processing after this is just carried out according to the identical order of step after the step S3 with above-mentioned Fig. 6.

As previously discussed, according to the data processing equipment of this variation, even at very big (or minimum) the absorbing wavelength λ not existing with target component _s0in the situation of relevant information, the sample of the standard specification that comprises this target component if can prepare, can carry out the peak value purity of the target component in unknown sample and judge.

[structure of the chromatography data system of the 2nd embodiment of the present invention and action]

Then,, with reference to Figure 12, the liquid chromatography system of the 2nd embodiment that possesses chromatography data system of the present invention is described.The principle of the 2nd embodiment based on above-mentioned explanation, by data processing, is separated in the retention time closer and by fully separated two target component x, y, do not undertaken quantitatively in cylinder 14.In Figure 12, the structure of LC portion 1 is identical with the structure of the 1st embodiment shown in Fig. 1, and therefore description thereof is omitted.

Identical with the 1st embodiment, data processing equipment 5 comprises: three-dimensional data storage portion 51, for preserving from the three-dimensional chromatographic data of A/D transducer 16 outputs; Wavelength chromatogram generating unit 52, according to the wavelength chromatogram of three-dimensional chromatographic data generation specific wavelength; Peak value test section 53 detects and carries out quantitative peak value, and sets its time range in this chromatogram; With two kinds of quantitative portions 54 of component separating, the peak value setting or time range separation on the basis by two target component x, y of operator's appointment, carry out respectively quantitatively.

Two kinds of quantitative portions 54 of component separating comprise as functional module: differential chromatogram generating unit 55, and very big (or minimum) absorbing wavelength λ x, λ y based on three-dimensional chromatographic data and two target component x, y, generate respectively differential chromatogram; Peak area calculating part 56, calculates the area of the peak value in present differential chromatogram; Quantitatively operational part 57, contrasts the area value and the lubber-line described later that calculate, obtains unknown target component x, the concentration of y; Lubber-line generating unit 58, the analysis result of the sample of target component x, y based on comprising concentration known, the peak area value in generation expression differential chromatogram and the lubber-line of the relation between constituent concentration; With lubber-line storage part 59, pre-stored generated lubber-line.

Distinctive data processing action in the liquid chromatography system of the 2nd embodiment is described.In the 2nd embodiment, for the concentration in unknown sample is that unknown target component x, y carries out respectively quantitatively, as following, generate in advance the lubber-line of target component x, y and be kept in advance in lubber-line storage part 59.

That is, operator dilutes the standard specification of target component x and modulates the standard model of multistage concentration, and the standard specification of similarly diluting target component y is modulated the standard model of multistage concentration.Then, by measuring respectively these standard models by LC portion 1, thereby obtain three-dimensional chromatographic data.The three-dimensional chromatographic data obtaining is temporarily stored in three-dimensional data storage portion 51.

Operator is by operating portion 4 input target component x, very big (or minimum) absorbing wavelength λ x of y, the wavelength value of λ y.Accept this input, the three-dimensional chromatographic data that two very big (or minimum) absorbing wavelength λ x, the λ ys of wavelength chromatogram generating unit 52 based on for inputted and each standard model obtain, generates respectively greatly very big (or minimum) absorbing wavelength chromatogram of (or minimum) absorbing wavelength λ x, λ y.Then, peak value test section 53 carrys out detection peak by the processing identical with the 1st embodiment, obtains the starting point T with each peak value _s, summit T ₀and terminal T _ethe corresponding time (now, origin only being detected in a peak value of target component x or y in very big (or minimum) absorbing wavelength chromatogram).

Differential chromatogram generating unit 55, for the standard model that comprises composition x with a certain concentration, is obtained the starting point T of the peak value relevant to composition x from three-dimensional data storage portion 51 _sto terminal T _etime range in absorbance spectrum, and for each absorbance spectrum, in wavelength direction, the absorbance of very big (or minimum) absorbing wavelength λ y of target component y is carried out to differential respectively, thereby obtains wavelength differential coefficient.Then, generate to have drawn and take the differential chromatogram of transverse axis as time, the longitudinal axis of take very big (or minimum) the absorbing wavelength λ y that is the wavelength differential coefficient that calculates.Now, because the standard model measuring only comprises composition x, therefore, in flowing out all time ranges of this composition x, under wavelength X x, absorbance represents greatly (or minimum) always.This is because greatly the differential coefficient of (or minimum) absorbing wavelength λ x remains 0.

With respect to this, although wavelength X y is not very big (or minimum) absorbing wavelength for composition x, also there is the absorption of composition x in wavelength X y.Therefore, the wavelength differential coefficient of very big (or minimum) absorbing wavelength λ y of composition y changes in the scope of absorption of accepting composition x., there is peak value in its result, this peak value reflects the outflow profile of composition x in the differential chromatogram of above-mentioned greatly (or minimum) absorbing wavelength λ y.The area value of the peak value of the composition x that therefore, peak area calculating part 56 calculating origin occur in the differential chromatogram at wavelength X y.In addition, the three-dimensional chromatographic data that the standard model of the composition x based on for comprising variable concentrations obtains, implements same calculating to the differential chromatogram of wavelength X y, calculates origin in the peak area value of composition x.

Peak area value and each constituent concentration of lubber-line generating unit 58 based on from obtaining the differential chromatogram of the wavelength X y corresponding with the standard model that comprises composition x with variable concentrations respectively as described above, generate and represent the concentration of composition x and the lubber-line of the relation between the peak area value of wavelength X y in differential chromatogram, and the relation obtaining is kept in lubber-line storage part 59.

In addition, similarly, the standard model of differential chromatogram generating unit 55 based on for comprising composition y and the three-dimensional chromatographic data that obtains, the differential chromatogram of very big (or minimum) absorbing wavelength λ x of generating component x, the area value of the peak value of the composition y that peak area calculating part 56 calculating origin occur in the differential chromatogram at wavelength X x.Then, the peak area value and each constituent concentration that the differential chromatogram of the wavelength X x of lubber-line generating unit 58 based on from corresponding with the standard model that comprises composition y with variable concentrations respectively, obtain, generate and represent the concentration of composition y and the lubber-line of the relation between the peak area value of wavelength X x in differential chromatogram, and the relation obtaining is kept in lubber-line storage part 59.

As previously discussed, can in lubber-line storage part 59, preserve composition x, y lubber-line separately.

Composition x, y in the sample of target component x, y to comprising concentration the unknown carries out when quantitative, by measuring this unknown sample by LC portion 1, thereby obtains three-dimensional chromatographic data and is kept in three-dimensional data storage portion 51.

Operator is by operating portion 4 input target component x, very big (or minimum) absorbing wavelength λ x of y, the wavelength value of λ y.Accept after this input, wavelength chromatogram generating unit 52 is read the three-dimensional chromatographic data of inputted two very big (or minimum) absorbing wavelength λ x, λ y from three-dimensional data storage portion 51, generate these greatly wavelength chromatograms of (or minimum) absorbing wavelength λ x, λ y.Then, peak value test section 53 is from two wavelength chromatograms, and the detection peak by processing same as the previously described embodiments, obtains the starting point T with each peak value _s, summit T ₀and terminal T _ethe corresponding time.Therefore but the peak value sometimes appearing in wavelength chromatogram can be two states that peak value is overlapping as Figure 15 (a), for example each composition x based on providing in advance, the retention time of y, using the starting point of the first half of connected peak value as peak value starting point T _s, using the terminal of latter half as peak value terminal T _ethereby, process.

In addition, identical with the 1st embodiment, by peak value, detecting a plurality of peak value detected in the situation that, on the picture of display part 3, showing the information of the peak value detecting, by operator, by operating portion 4, among these a plurality of peak values, selecting origin in the target peak of target component.

Then, differential chromatogram generating unit 55 is obtained the absorbance spectrum in the time range of or origin-to-destination that selected by operator, peak value detected by peak value test section 53 from three-dimensional data storage portion 51, respectively to each absorbance spectrum, in wavelength direction, the absorbance of very big (or minimum) absorbing wavelength λ y of very big (or minimum) absorbing wavelength λ x of target component x and target component y is carried out to differential, thereby obtain wavelength differential coefficient.Then, generate respectively the differential chromatogram of very big (or minimum) absorbing wavelength λ x and λ y.As mentioned above, the differential chromatogram of wavelength X x can not show the impact of the absorption of target component x, and the peak value observing has reflected the outflow profile of target component y.On the other hand, the differential chromatogram of wavelength X y can not show the impact of the absorption of target component y, and the peak value observing has reflected the outflow profile of target component x.Therefore, peak area calculating part 56 calculates respectively the area value of the peak value occurring in each differential chromatogram.

Quantitatively operational part 57 contrasts the lubber-line of the peak area value of obtaining according to the differential chromatogram of wavelength X x and the composition y reading from lubber-line storage part 59, thereby calculates the concentration value of composition y.In addition, contrast the lubber-line of the peak area value of obtaining according to the differential chromatogram of wavelength X y and the composition x reading from lubber-line storage part 59 equally, thereby calculate the concentration value of composition x.Then, the quantitative result of two target component x, y that obtain like this to operator notification by display part 3.

As previously discussed, in the relevant data processing equipment 5 of the 2nd embodiment, according to the area value of the peak value occurring in the differential chromatogram of very big (or minimum) absorbing wavelength of other compositions different from target component, target component is carried out quantitatively after having removed the impact of this composition of overlapping outflow.Now, appear at the outflow profile that peak value in differential chromatogram has only reflected target component, therefore, with the carrying out quantitative method by vertical segmentation after to overlapping the cutting apart of peak value and compare of prior art, can obtain more high-precision quantitative result.

In the 2nd embodiment, also can realize illustrated various distortion in above-mentioned the 1st embodiment.For example, except being inputted the wavelength value of very big (or minimum) absorbing wavelength λ x, λ y of target component x, y by operator, can also, by the title of operator's intended target composition or structural formula etc., also can from database, obtain wavelength value corresponding thereto.From this database, can also side by side obtain the retention time with the wavelength value of any composition, be particularly advantageous in the situation that of therefore utilizing the retention time when carrying out peak value detection.

In addition, the present invention is not limited to above-mentioned the 1st embodiment, the 2nd embodiment, has carried out suitable distortion, appends, revises in the scope that is also obviously included in the application's claims within the scope of aim of the present invention.

For example, the chromatographic detecting device that data processing equipment of the present invention obtains the three-dimensional chromatographic data of processing object can not be the multi-channel type detecting devices such as PDA detecting device, as long as according to obtaining the correct mode that has reflected the differential coefficient of the inclination of its curve of spectrum while successively the absorbance of absorbance spectrum having been carried out to differential in wavelength direction, obtain the spectrum of waveform shape wider (changing slowly).But, absorbance measuring spended time within the scope of whole provision wavelengths is long is unaccommodated, can be therefore the ultra-violet and visible spectrophotometer that can carry out quick length scanning, infrared light spectrophotometer, near infrared light spectrophotometer, fluorospectrophotometer etc.

In addition, chromatography can not be liquid chromatography, but gas chromatography, and the chromatography of using detecting device as above is liquid chromatography normally.In addition, as mentioned above, can clearly can also in the device that following data are processed or method, apply the present invention, these data are not to detect by detecting device the data that obtained by the sample of chromatographic cylinder separation, but are detected without carrying out by FIA method the data that component separating obtains with regard to the composition in the sample being imported into by detecting device.

Symbol description

1 ... LC portion

11 ... mobile phase container

12 ... liquid-feeding pump

13 ... sample injection unit

14 ... cylinder

15 ... PDA detecting device

16 ... A/D transducer

2,5 ... data processing equipment

21,51 ... three-dimensional data storage portion

22,52 ... wavelength chromatogram generating unit

23,53 ... peak value test section

24 ... impurity test section

25 ... differential chromatogram generating unit

26 ... detection unit

27 ... very big (or minimum) absorbing wavelength acquisition unit

3 ... display part

4 ... operating portion

54 ... two kinds of quantitative portions of component separating

55 ... differential chromatogram generating unit

56 ... peak area calculating part

57 ... quantitative operational part

58 ... lubber-line generating unit

59 ... lubber-line storage part

Claims (10)

1. a chromatography data system, processes the three-dimensional chromatographic data that time, wavelength and absorbance be dimension of take of collecting for target sample, and this chromatography data system is characterised in that to possess:

A) differential chromatogram generation unit, based on described three-dimensional chromatographic data, for the wavelength of each time point in all time ranges of expression or within the scope of the stipulated time and the absorbance spectrum of the relation of absorbance, obtain differential coefficient in wavelength direction of the very big absorbing wavelength of the 1st composition or minimum absorbing wavelength, be wavelength differential coefficient, generate the differential chromatogram that represents that time of the described wavelength differential coefficient in all time ranges or within the scope of the stipulated time changes; With

B) chromatogram waveform processing unit, the waveform based on described differential chromatogram, determines whether quantitative to the 2nd overlapping composition of the peak value of Multiple components or execution and described the 1st composition of overlapping other of the peak value that exists with described the 1st composition 1.

2. chromatography data system according to claim 1, is characterized in that,

Described chromatography data system also possesses: c) wavelength chromatogram generation unit, and based on described three-dimensional chromatographic data, about the absorbing wavelength of the 1st composition and generate the wavelength chromatogram of the relation between expression time and absorbance,

Described differential chromatogram generation unit is following unit: based on described three-dimensional chromatographic data, the absorbance spectrum of each time point for the peak value that comprises the target component in described wavelength chromatogram in interior time range, obtain the wavelength differential coefficient of very big absorbing wavelength or the minimum absorbing wavelength of described the 1st composition, generate the differential chromatogram of the time variation that represents this wavelength differential coefficient

Described chromatogram waveform processing unit is following identifying unit: whether the waveform shape based on described differential chromatogram, comprise impurity in the peak value of judgement as described the 1st composition of target component.

3. chromatography data system according to claim 2, is characterized in that,

Whether described identifying unit is smooth by judging described differential chromatogram, thereby judges whether the peak value of target component comprises impurity.

4. according to the chromatography data system described in claim 2 or 3, it is characterized in that,

Described chromatography data system also possesses: d) peak detection unit, detect the peak value of described wavelength chromatogram, and determine starting point and the terminal of peak value,

Described differential chromatogram generation unit generates the differential chromatogram in the time range of origin-to-destination of peak value of the target component in described wavelength chromatogram.

5. according to the chromatography data system described in any one of claim 2～4, it is characterized in that,

Described chromatography data system also possesses: e) very big absorbing wavelength acquiring unit or minimum absorbing wavelength acquiring unit, the summit of the peak value of the wavelength chromatogram of the three-dimensional chromatographic data in wavelength direction, the sample based on for comprising known target composition being obtained absorbance spectrum is constantly carried out differential, thereby obtains very big absorbing wavelength or the minimum absorbing wavelength of described target component.

6. chromatography data system according to claim 1, is characterized in that,

Described differential chromatogram generation unit is based on described three-dimensional chromatographic data, absorbance spectrum for each time point in all time ranges or within the scope of the stipulated time, obtain the very big absorbing wavelength of described the 1st composition or the wavelength differential coefficient of minimum absorbing wavelength as the 1st target component, generate the differential chromatogram of the time variation that represents this wavelength differential coefficient, and obtain the very big absorbing wavelength of described the 2nd composition or the wavelength differential coefficient of minimum absorbing wavelength as the 2nd target component, generate the differential chromatogram of the time variation that represents this wavelength differential coefficient

The peak value occurring in the differential chromatogram of the very big absorbing wavelength of described chromatogram waveform processing unit based at described the 1st target component or minimum absorbing wavelength carries out quantitatively described the 2nd target component, and the peak value occurring in the differential chromatogram of the very big absorbing wavelength based at described the 2nd target component or minimum absorbing wavelength carries out quantitatively described the 1st target component.

7. chromatography data system according to claim 6, is characterized in that,

Described chromatogram waveform processing unit comprises:

Inspection amount information memory cell, for described the 1st target component and the 2nd target component, the inspection amount information of the relation between the pre-stored area that is illustrated in the peak value occurring in differential chromatogram or height and constituent concentration respectively;

Peak information computing unit, calculates the three-dimensional chromatographic data based on for target sample and the area or the height that generate respectively, appear at the peak value in differential chromatogram in the very big absorbing wavelength of described the 1st target component or the very big absorbing wavelength of minimum absorbing wavelength and described the 2nd target component or minimum absorbing wavelength; With

Quantitative values computing unit, contrasts the area of the peak value being calculated by this peak information computing unit or height with described inspection amount information, obtain the quantitative values of each target component.

8. a chromatographic data disposal route, processes the three-dimensional chromatographic data that time, wavelength and absorbance be dimension of take of collecting for target sample, and this chromatographic data disposal route is characterised in that, comprising:

A) differential chromatogram generates step, based on described three-dimensional chromatographic data, for the wavelength of each time point and the absorbance spectrum of the relation between absorbance that represent in all time ranges or within the scope of the stipulated time, obtain differential coefficient in wavelength direction of the very big absorbing wavelength of the 1st composition or minimum absorbing wavelength, be wavelength differential coefficient, generate the differential chromatogram that represents that time of the described wavelength differential coefficient in all time ranges or within the scope of the stipulated time changes; With

B) chromatogram waveform processing step, the waveform based on described differential chromatogram, determines whether quantitative to the 2nd overlapping composition of the peak value of Multiple components or execution and described the 1st composition of overlapping other of the peak value that exists with described the 1st composition 1.

9. chromatographic data disposal route according to claim 8, is characterized in that,

Also comprise: c) wavelength chromatogram generates step, based on described three-dimensional chromatographic data, about the absorbing wavelength of the 1st composition, generate the wavelength chromatogram of the relation of expression time and absorbance,

In described differential chromatogram, generate in step, based on described three-dimensional chromatographic data, the absorbance spectrum of each time point for the peak value that comprises the target component in described wavelength chromatogram in interior time range, obtain the wavelength differential coefficient of very big absorbing wavelength or the minimum absorbing wavelength of described the 1st composition, generate the differential chromatogram of the time variation that represents this wavelength differential coefficient

In described chromatogram waveform processing step, whether the waveform shape based on described differential chromatogram, comprise impurity in the peak value of judgement as described the 1st composition of target component.

10. chromatographic data disposal route according to claim 8, is characterized in that,

In described differential chromatogram, generate in step, based on described three-dimensional chromatographic data, absorbance spectrum for each time point in all time ranges or within the scope of the stipulated time, obtain the very big absorbing wavelength of described the 1st composition or the wavelength differential coefficient of minimum absorbing wavelength as the 1st target component, generate the differential chromatogram of the time variation that represents this wavelength differential coefficient, and obtain the very big absorbing wavelength of described the 2nd composition or the wavelength differential coefficient of minimum absorbing wavelength as the 2nd target component, generate the differential chromatogram of the time variation that represents this wavelength differential coefficient,

In described chromatogram waveform processing step, the peak value occurring in very big absorbing wavelength based at described the 1st target component or the differential chromatogram of minimum absorbing wavelength carries out quantitatively described the 2nd target component, and the peak value occurring in the differential chromatogram of the very big absorbing wavelength based at described the 2nd target component or minimum absorbing wavelength carries out quantitatively described the 1st target component.

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